Engineering yeast with a light-driven proton pump system in the vacuolar membrane

Area of Science:

• Synthetic biology
• Metabolic engineering
• Biotechnology

Background:

• Cellular metabolism relies on a sufficient supply of adenosine triphosphate (ATP).
• Efficient bioproduction in cell factories is constrained by ATP availability.
• Vacuolar acidification by V-ATPase consumes significant cellular ATP.

Purpose of the Study:

• To engineer yeast with a light-driven proton pump in the vacuolar membrane.
• To reduce ATP consumption by V-ATPase.
• To increase intracellular ATP supply for enhanced bioproduction.

Main Methods:

• Constructed a light-driven proton pump using Delta rhodopsin (dR) from Haloterrigena turkmenica.
• Localized dR to the vacuolar membrane of Saccharomyces cerevisiae.
• Evaluated light-driven proton pumping activity and measured intracellular ATP content.

Main Results:

• Successfully expressed and localized dR in the yeast vacuolar membrane.
• Demonstrated light-driven proton pumping activity in isolated vacuoles.
• Observed a light-induced increase in intracellular ATP content in engineered yeast.

Conclusions:

• Engineered yeast with a vacuolar light-driven proton pump can serve as an optoenergetic cell factory.
• This approach offers a novel strategy to enhance ATP supply for bioproduction.
• Potential applications in various bioproduction processes requiring efficient energy metabolism.